Anti-Human IgG3 Antibody [KT133]

Various protein samples were run on 6-18% SDS-PAGE under reducing conditions and blotted onto nitrocellulose membrane. KT133 at 1 µg/mL was used as the primary antibody and peroxidase conjugated goat anti-mouse IgG was used as the secondary antibody. Human IgG3 band was visualized using ECL Western Blotting Substrate. 

Lane 1: 100 ng of rat IgG1 

Lane 2: 100 ng of human IgE 

Lane 3: 100 ng of human IgM 

Lane 4: 100 ng of human IgA 

Lane 5: 100 ng of human IgG4 

Lane 6: 100 ng of human IgG3 

Lane 7: 100 ng of human IgG2 

Lane 8: 100 ng of human IgG1 

Result: KT133 can specifically detect human IgG3 by Western blotting.

Microtiter wells were coated with KT133 at 3 µg/mL as the capture antibody. Human IgG3 was used as the antigen. Peroxidase conjugated mouse anti-human IgG3 monoclonal antibody (K06089_8E4) was used as the detection antibody. 

Result: KT133 and K06089_8E4 can be used as a matched antibody pair to detect and quantify the concentration of human IgG3.

IHC-P analysis of human tonsil tissue by anti-human IgG3 antibody (KT133). 

IHC-P was performed using sections of the formalin-fixed paraffin-embedded human tonsil tissue. Antigen was retrieved through addition of boiling Tris/EDTA buffer pH 9 in a pressure cooker for 3 min. Endogenous peroxidase activity was quenched by incubating the sections with 3% H2O2 for 30 min at room temperature. The sections were then incubated with anti-human IgG3 primary antibody (KT133) at 1 µg/mL at room temperature for 1 h. Poly-peroxidase conjugated goat anti-mouse IgG was used as the secondary antibody. Diaminobenzidine was used as the chromogen. The section was counterstained with hematoxylin. 

Result: Germinal center cells and squamous epithelial cells are positively stained at cytoplasm and membranous.

Microtiter wells were coated with human immunoglobulins with various isotypes. KT133 was used as primary antibody and peroxidase conjugated goat anti-mouse IgG was used as the secondary antibody. 

Result: KT133 does not cross-react with human IgG1, IgG2, IgG4, IgA, IgM and IgE.